Fluorescence Microscopy Shared Resource Instrumentation

  1. Zeiss LSM 800 AiryScan Confocal Microscope
  2. Leica TCS-SP8 Confocal Microscope & FLIM System
  3. Zeiss AxioObserver Microscope with Hamamatsu Flash4.0v2 sCMOS Camera
  4. Olympus DSU Spinning Disk Confocal/Stereology System
  5. Olympus IX71 microscope with Andor iXon 897 EMCCD Camera
  6. Zeiss AxioPlan2 Microscope with Nuance Multi-Spectral Camera
  7. Zeiss Axioskop Microscopewith AxioCamHR Color Camera
  8. Nikon TE2000 Microscope with Q-Imaging Retiga R3 Camera (monochrome)
  9. Image Analysis Workstations
  10. GE Typhoon FLA 7000 Scanner
  11. Li-Cor Odessey FC


  1. Zeiss LSM 800 AiryScan Confocal Microscope
    Location: CRF226

    Microscope Details

    • Mounted on Inverted (AxioObserver) microscope with a motorized X-Y stage
    • Metal Halide fluorescence and LED transmission lamp.
    • Solid state diode laser excitation (405, 488, 561 & 640) for DAPI, FITC, Rhodamine, & Cy5-like fluorophores
    • High sensitivity GaAsP PMT detectors (2) for conventional imaging and a single AiryScan detector for high resolution imaging near super-resolution capabilities (~140-150nm)
    • Transmitted light detector for acquiring simultaneous DIC images
    • Objectives: 10x, 20x, 40x oil DIC, 63x oil DIC (all fluorescence objectives)
    • Zen 2 Imaging software for conventional confocal mode, AiryScan, and spectral acquisition for "unmixing" overlapping fluorophores or autofluorescence.
    • Multi-track (sequential) image acquisition reduces channel cross-talk in multiple labeling experiments

    Possible Applications

    • Fixed specimen mounted on slides
    • Live cells in coverslip dish/heating with full microscope enclosure
    • Confocal Fluorescence – specimens labeled with DAPI, CFP, YFP, FITC, Rhodamine, Cy5-like fluorophores
    • Near super-resolution imaging with AiryScan (63X objectives only) and post hoc image processing.
    • Spectral imaging and unmixing for up to 10 fluorophores; for autofluorescence removal and using fluorophores with overlapping emission spectra
    • DIC (non-confocal) with 40X and 63X objectives, merged with fluorescence or alone
    • FRAP (fluorescence recovery after photobleaching)
    • FRET (fluorescence resonance energy transfer)
    • Acquires a series of images along the Z axis (Z-stack) and processes the images to produce a 3-D projection image
    • Multi-position and tile scanning with use of the X-Y motorized stage
    • Live-cell (time lapse) imaging capabilities with Pecon stage top incubation system and full enclosure for long term environmental (CO2 and temperature) stability.
    • Two sample holder inserts for cells grown on 25 mm diameter glass coverslips or multi-well cover slip bottom chambers.
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  3. Leica TCS-SP8 Confocal Microscope & FLIM System
    Location: CRF216

    Microscope Details

    • DMI8 inverted microscope
    • Metal halide fluorescence lamp & halogen transmission lamp
    • 10X/0.4NA, 20X/0.75NA multi-immersion (oil, water, glycerol), 25X/0.95 NA LD water, 63X/1.4NA oil, & 63X/1.2NA water objectives
    • DIC available with 63X oil and 63X water objectives
    • Three lasers: tunable, pulsed white light laser (470-670nm); 405 nm diode laser; 5-line argon laser (452, 476, 488, 496, 514 nm)
    • Laser excitation for most fluorophores (405 and 470-670nm excitation)
    • Five fluorescence detectors: 2 sensitive hybrids (Leica HyDs); 2 HyD SMD (cooled hybrids for single molecule detection); 1 standard PMT.
    • Prism-based emission fluorescence separation providing maximum flexibility in specifying emission wavelength ranges.
    • Motorized XY stage.
    • Two Z axis control modes:  standard motorized focus control plus Z galvo stage for piezo-driven positioning of slide/sample chamber.
    • LASX image acquisition and analysis software
    • Huygens Essential software for deconvolution (deblurring) of Z stacks

    Possible Applications

    • Fixed specimen mounted on slides
    • Live cells in coverslip dish/heating with objective heater (full stage incubation system for controlling CO2, temperature, and humidity, made available in 2016)
    • Confocal Fluorescence – specimens labeled with most fluorophores
    • Spectral imaging and unmixing for up to 10 fluorophores; for autofluorescence removal and using fluorophores with overlapping emission spectra
    • Excitation spectrum can be collected with white light laser (470-670 nm)
    • DIC (non-confocal) with 63X objectives, merged with fluorescence or alone
    • Z-stacks, time series, and montage imaging
    • Multi-position and tile scanning with use of the X-Y motorized stage
    • Quantitative fluorescence measurements using HyD detectors in photon counting mode
    • FLIM (fluorescence lifetime imaging)
    • FRAP (fluorescence recovery after photobleaching)
    • FRET (fluorescence resonance energy transfer)
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  5. Zeiss AxioObserver Microscope with Hamamatsu Flash4.0v2 sCMOS Camera
    Location: CRF212

    Microscope Details

    • Zeiss AxioObserver inverted fluorescence microscope
    • Hamamatsu ORCA Flash4.0v2 sCMOS monochrome camera (100fps at full resolution 2048x2048)
    • Metal halide fluorescent lamp & halogen transmission lamp
    • 10x, 20x, 63xDIC Oil, & 100x Oil objectives
    • Phase Contrast for 10x and 20x objectives
    • Fluorescence filter sets include: DAPI, FITC, Rhodamine/TRITC/Cy3, Texas Red & Cy5
    • DIC available with 63x objectives
    • Objective heater (63x water objective)
    • Motorized nosepiece for Z-stacking
    • Motorized X-Y stage for multi-position image acquisition (fixed or live-cell time lapse imaging)
    • SlideBook6 image acquisition software

    Possible Applications

    • Fixed specimen mounted on slides
    • Live cells in coverslip dish/heating with objective heater
    • DIC (non-confocal) with 63x objectives, merged with fluorescence or alone
    • Z-stacks, montage,andtime series imaging
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  7. Olympus DSU Spinning Disk Confocal/Stereology System
    Location: CRF212

    Microscope Details

    • Olympus IX-81 inverted microscope
    • Metal halide fluorescence lamp and halogen transmission lamp
    • 4x, 10x, 20x, 20x Oil, 40x, 60x Oil DIC, 60x Water DIC, 100x Oil objectives
    • X,Y, Z motorized stage
    • Pathology Devices live cell incubation system w/ CO2, heat, humidity
    • Color digital camera
    • EM CCD high sensitivity monochrome camera
    • Stereo Investigator software for design based stereology
    • Neurolucida software for neuron tracing

    Possible Applications

    • Fixed specimen mounted on slides
    • Live cell imaging
    • Confocal and widefield fluorescence
    • Stereology
    • X,Y positioning, Z-stacks, time series, and montage imaging
    • DIC with both 60x objectives
    • Brightfield – H&E, DAB, Giemsa, etc
    • Darkfield
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  9. Olympus IX71 microscope with Andor iXon 897 EMCCD Camera
    Location: CRF218

    Microscope Details

    • Inverted Olympus IX-71 inverted microscope
    • Mercury fluorescence lamp and halogen transmission lamp
    • Monochromator – 350nm-670nm excitation range
    • Lasers: 405nm, 488nm,520NM and 642nm
    • 20x, 60x H2O, 100x Oil, 150x Oil TIRF objectives
    • Fluorescence filters for Quantum Dots, DAPI, FITC, & Rhod-like fluorophores, Fura-2, Indo-1
    • Andor IQ Image Acquisition software
    • Objective heater and Pathology Devices live cell incubation system w/ CO2, heat, humidity
    • XYZ motorized stage
    • Andor iXon EM-CCD camera
    • Opti-split (dual) Image splitter

    Possible Applications

    • Fixed specimen mounted on slides
    • Live cell imaging with objective warmer
    • Single Particle Tracking
    • Total Internal Reflection Fluorescence (TIRF)
    • Ratio-metric Imaging (dual excitation/single emission or single excitation dual emission)
    • Live cells in culture
    • Widefield Fluorescence – many fluorophores possible with monochromator
    • Z-stacks & time series
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  11. Zeiss AxioPlan2 Microscope with Nuance Multi-Spectral Camera
    Location: CRF224

    Microscope Details

    • AxioPlanMOT upright microscope
    • Metal Halide fluorescence lamp & halogen transmission lamp
    • 10x, 20x, 40x Oil & 63x Oil objectives
    • DAPI, FITC, Rhodamine, Cy5 long pass filters for use with Nuance camera.
    • Nuance spectral camera and software
    • Motorized nosepiece, Z-stage, filter turret & shutter for easy operation.

    Possible Applications

    • Fixed specimen mounted on slides
    • Widefield fluorescence – specimen labeled with DAPI, FITC, Rhod, & Cy5-like fluorophores
    • Brightfield – H&E, DAB, Giemsa, etc.
    • Nuance spectral camera can be used to separate closely overlapping fluorophores, histochemical stains, distinguish autofluorescence and quantify labeling
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  13. Zeiss Axioskop Microscope with AxioCam HR Color Camera
    Location: CRF226

    Microscope Details

    • AxioSkop 2 MOT upright microscope
    • 4x, 10x, 20x, 40x, 40x Oil & 63x Oil
    • Phase Contrast for 10x and 20x objectives
    • AxioCam HR digital color camera
    • Zeiss AxioVision image acquisition software
    • Motorized Z-stage

    Possible Applications

    • Fixed specimen mounted on slides for general histopathology
    • Brightfield – H&E, DAB, Giemsa, etc.
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  15. Nikon TE2000 Microscope with Q-Imaging Retiga R3 Camera (monochrome)
    Location: CRF216

    Microscope Details

    • Nikon TE2000 inverted microscope
    • XCITE120LED illumination source for fluorescence applications. Ideal for hit and run quality control in tissue culture fluorescence constructs
    • Halogen transmission lamp
    • 10x, 20xDIC, 40xDIC Oil, 60xDIC Oil objectives
    • DIC available with all objectives except the 10x
    • Fluorescence filters for DAPI, CFP, YFP, FITC, Rhodamine-like & Cy5 fluorophores
    • Q-Imaging Retiga R3 monochrome camera and Ocular image acquisition software for basic image documentation.

    Possible Applications

    • Fixed specimen mounted on slides
    • Live cells in culture – dishes, plates, and flasks.
    • Widefield Fluorescence – specimen labeled with DAPI, CFP, YFP, FITC, Rhod-like & Cy5 dyes
    • DIC 20-60X, merged with fluorescence or alone
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  17. Image Analysis Workstations

    Locations: CRF224 & CRF226

    Workstation Details

    Software

    • Huygens Deconvolution Scientific Volume Imaging (1)
    • SlideBook (1)
    • StereoInvestigator (1)
    • Neurolucida (1)
    • LASX (Leica confocal) (1)
    • IQTL (Typhoon scanner analysis) (1)
    • Zen (2)
    • LSM510 (2)
    • Nuance spectral imaging software (2)

    Analysis Options

    • Co-localization
    • Deconvolution
    • Spectral unmixing
    • Intensity analysis
    • Morphological analysis
    • 3D rendering
    • Object counting
    • Particle Tracking
    • Preparation for publication
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  19. GE Typhoon FLA 7000 Scanner

    Locations: CRF214

    Scanner Details: Typhoon FLA 7000 is a fast and versatile laser scanner for biomolecular imaging applications including sensitive and quantitative measurements of radioisotopic labels, chemifluorescent Western blots, and single fluorescence (visible excitation), as well as documentation of colorimetric stains (e.g., Coomassie blue and silver-stained gels).

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  21. Li-Cor Odessey FC

    Locations: CRF214

    Scanner Details: Li-Cor Odessey Fc is a near infra-red scanner (680nm & 740nm) for biomolecular imaging applications including sensitive and quantitative measurements of chemiluminescent Western blots as well as documentation of colorimetric stains (e.g., Coomassie blue and silver-stained gels).

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